QIAamp DNA FFPE Tissue Kit

For purification of genomic DNA from formalin-fixed paraffin-embedded tissues

Rapid purification of high-quality, ready-to-use DNA

Consistent, high yields

Complete removal of contaminants and inhibitors

The QIAamp DNA FFPE Tissue Kit is specially designed for purifying DNA from formalin-fixed, paraffin-embedded tissue sections. The kit uses special lysis conditions to release DNA from tissue sections and to overcome inhibitory effects caused by formalin crosslinking of nucleic acids. The kit uses QIAamp MinElute spin columns for purification of high-quality DNA in small volumes. Purification of DNA using the QIAamp DNA FFPE Tissue Kit can be automated on the QIAcube.

Genomic DNA was purified from different FFPE tissue samples using the QIAamp DNA FFPE Tissue Kit. Products of the prnp gene (78 bp, 464 bp, and 727 bp) were amplified by real-time quantitative PCR.|Paraffin is dissolved in xylene and removed. The sample is lysed under denaturing conditions with a short proteinase K digestion. Incubation at 90°C reverses formalin crosslinking. DNA binds to the membrane and contaminants flow through. Residual contaminants are washed away. DNA is eluted in Buffer ATE or water, and is immediately ready for use in amplification reactions or for storage at –20°C. Purified DNA is free of proteins, nucleases, and other impurities.|

The QIAamp DNA FFPE Tissue Kit purifies DNA that can be used in a wide range of downstream applications, such as single nucleotide polymorphism (SNP) and short-tandem repeat (STR) genotyping and pharmacogenomic research. DNA purified with the QIAamp DNA FFPE Tissue Kit can also be used in PCR (see figure "PCR analysis of DNA purified from FFPE tissue samples").

Principle

The QIAamp DNA FFPE Tissue Kit uses well-established QIAamp MinElute technology for purification of genomic and mitochondrial DNA from small sample volumes or sizes. The kit combines the selective binding properties of a silica-based membrane with flexible elution volumes of between 20 and 100 μl.

Specially optimized lysis conditions allow genomic DNA to be efficiently purified from FFPE tissue sections without the need for overnight incubation. Incubation at an elevated temperature after proteinase K digestion partially removes formalin crosslinking of the released DNA, improving yield as well as DNA performance in downstream assays. Note that DNA isolated from FFPE samples is usually of lower molecular weight than DNA from fresh or frozen samples. The degree of fragmentation depends on the type and age of the sample and the conditions used for fixation.